Analysis of Lipids in Egg Yolk

Rojas, Kristine Loren P. Roldan, Hyvie Joyce L.

Sampang, Juvenis O. Senseng, Alyssa April L.
Group 6 – 2KPHC

ABSTRACT

In this experiment we have extracted the lipids in an egg yolk and perform a qualitative
test for lipids. The tests performed were the Test for Ester, Test for Choline also known as the
Kraut’s Test and the Test for Cholesterol also known as the Liebermann-Buchard Test. In the
separated egg yolk a Hexane: Ethanol mixture was added and have stand for about 5 minutes.
The upper part of the extract was collected and a pinch of Na2SO4 was added. The extract was
used in the different tests. The lipid extracted was a yellow oily solution. The result have showed
that the lipid extract of the yolk of the egg has an ester and a cholesterol, and the egg yolk extract
showed a negative result in the test for choline, for it doesn’t have any change in color.

INTRODUCTION
Lipids are broadly defined as any fat-
soluble (lipophilic), naturally-occuring
molecule, such as fats, oils, waxes,
cholesterol, sterols and fat soluble vitamins
(such as vitamins A, D, E and K). Biological
functions of lipids include energy storage,
acting as structural components of cell
membranes, and participating as important
signaling molecules.
Lipids are insoluble in water but
soluble in non-polar solvents and solvents of
low polarity. This lack of solubility in water
is a very important property since our body
chemistry is firmly based on water.
In this experiment, the group gained
familiarity with one of the separation
methods used in lipid biochemistry:
extraction.
The general approach is to extract lipids
from egg yolk using hexane-ethanol mixture
(2:1) twice the volume of the egg yolk. The
extracts were collected and a pinch of sodium
sulfate was added and the stored in an amber
bottle.
METHODOLOGY
A. Extraction of Lipids in Egg yolk
The group used 5 eggs and
separated the egg yolks from the
white. The volume of the yolks
(yellow-colored) was determined
using a graduated cylinder. After
determining the volume, the yolks
were transferred on a beaker and
hexane-ethanol mixture was added
twice the volume on the said yolks.
The solution was mixed and allowed
to stand for 5 minutes. Two separate
layers was produced. A pinch of
sodium sulfate was added on the
solution. After a while, the upper
layer (liquid portion) of the solution
was extracted and put in an amber
bottle.
B. Qualitative Test for Lipids
1.) Test for Ester:
Ten drops of extract was placed on a test tube
and 0.5 mL of ethanol:1-butanol together
with two drops each of 2M NH2OH HCl and
3M of NaOH was added. The solution was
mixed well and allowed to stand for 5
minutes. Afterwards, 2 drops of the 6M HCl
and a drop of the 5% FeCl3 ⋅6H2O was added
in the mixture.

2.) Test for Choline

Three mL of Kraut’s reagent was placed on a
test tube containing 10 drops of the extract.
The test tube was warmed and observed
afterwards.
Figure 1. The results of the tests performed.
3.) Test for Cholesterol
While in the test for Choline the egg
Ten drops of the extract was placed in the test
yolk lipid is negative where there is no
tube and 0.25mL of dcm was added together
change of color nor formation of precipitate.
with 6 drops of acetic anhydride and 2 drops
of concentrated H2SO4 .
.
RESULTS AND DISCUSSION CONCLUSION AND
RECOMMENDATION
Table 1: Qualitative Test for Lipids Results
On the qualitative test for lipids, the
ESTER CHOLINE CHOLESTEROL egg yolk is positive to the Ester test and Test
EGG (+) (-) (+) for Cholesterol and is negative with the
YOLK Burg No Greenish Choline test.
undy Change Solution
Soluti REFERENCES
on
Ganong,B. (2007), Qualitative Test for
The test for Ester the Carboxylate Lipids
esters are converted to hydroxamic acids http://faculty.mansfield.edu/bganong/bioche
which form a burgundy colored complex mistry/lipids3.htm
with ferric ion, that shows a positive result.
Introduction to general, organic, and
For the test of Cholesterol or the Biochemistry. Bettelheim, brown, Campbell
Liebermann-Buchard Test the greenish color and Farell Page, 528-534
is due to the hydroxyl group (-OH) of the
cholesterol reacting with the reagents and
increasing the conjugation of the un-
saturation in the adjacent fused ring resulting
to a positive result.